Abstract:
:A multiplex real-time PCR assay was developed to detect and quantify the Citrus tristeza virus (CTV) genotypic mixture present in infected plants. CTV isolate FS627, a complex Florida isolate containing T36, T30 and VT genotypes and its aphid transmitted subisolates was used. The relative quantitative assay was carried out using specific primers and probes developed from the genotypes of three CTV virus isolates and included the coat protein region of isolate T36 and the 5' end, ORF 1a and ORF 2 region of isolates T36, T30 and VT. Among the three genotypes present in the aphid transmitted subisolates, the T30 genotype showed higher overall relative quantitation in all specific regions compared to other isolates. The profiles of the some aphid transmitted subisolates were different from the parent source from which they transmitted. The 2(-DeltaDeltaCt) method (the amount of target, normalized to an endogenous control and relative to a calibrator) was used to analyze the relative titers of the three reference genotypes in the aphid transmitted plants infected with FS627. This protocol enabled assessments of CTV genetic diversity in the aphid transmitted subisolates. This simple quantitative assay was sensitive, efficient, and took less time than other existing methods. This relative quantitative assay will be a reliable tool for diagnosis, detection and genetic diversity studies on CTV.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Ananthakrishnan G,Venkataprasanna T,Roy A,Brlansky RHdoi
10.1016/j.jviromet.2009.12.001subject
Has Abstractpub_date
2010-03-01 00:00:00pages
75-82issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(09)00519-9journal_volume
164pub_type
杂志文章abstract::A sensitive and specific blocking enzyme-linked immunosorbent assay (ELISA) was developed to distinguish infectious bovine rhinotracheitis virus (IBRV)-infected animals from those immunized with a glycoprotein gIII deletion mutant, IBRV(NG)dltkdlgIII. For this ELISA, undiluted test sera are used to block the binding o...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(92)90006-y
更新日期:1992-10-01 00:00:00
abstract::A cell line which can be used in a simple, sensitive, and rapid histochemical assay was isolated for detection of herpes simplex virus (HSV). The cell line was derived by selection of G418 resistant colonies following co-transfection of baby hamster kidney cells with a plasmid which contains a G418 antibiotic resistan...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(92)90110-y
更新日期:1992-08-01 00:00:00
abstract::Accurate detection and quantitation of viruses can be beneficial to plant-virus interaction studies. In this study, three SYBR green real-time RT-PCR assays were developed to quantitate grapevine leafroll-associated virus 3 (GLRaV-3) in infected vines. Three genomic regions (ORF1a, coat protein and 3'UTR) were targete...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.09.022
更新日期:2014-12-15 00:00:00
abstract::The reverse transcriptase of the sheep lentivirus visna/maedi virus has been characterised. Optima for magnesium ion concentration (5-10 mM), potassium ion concentration (150 mM) and pH (8.25) for this enzyme are very similar to those previously described for the human immunodeficiency viruses. The assay used for this...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(92)90144-3
更新日期:1992-02-01 00:00:00
abstract::Specific cell-surface binding is the essential first step for cellular invasion by viruses. To understand this process, various methods to evaluate binding properties of viruses to cells have been developed. However, many rely on radioactive labeling or indirect immunofluorescence. The development of a novel fluoresce...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.08.011
更新日期:2006-06-01 00:00:00
abstract::A novel multiplex nested PCR (nPCR) method was developed for detecting and differentiating simultaneously the DNA of polyomaviruses JC, BK and SV40 in a single tube. In the first amplification step the same set of primers were used to amplify a conserved DNA region of the large T antigen gene of JCV, BKV and SV40. The...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(99)00095-6
更新日期:1999-10-01 00:00:00
abstract::The technique of gold-labelled immunosorbent electron microscopy for the initial screening of monoclonal antibodies 10 days after cell fusion from 96-well culture plates is described. The technique is used to identify clones that secrete antibodies binding on the surface of the virion or to viral subunits, and compare...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(88)90118-8
更新日期:1988-12-01 00:00:00
abstract::Specific domains of the NS4 and NS5 gene products of hepatitis C virus have been identified using hydrophilicity profiles for the prediction of potential immunogenic regions, and epitope scanning techniques. Peptides synthesised on the basis of such data show excellent reactivity in the ELISA format. Introduction of a...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(95)00060-8
更新日期:1995-10-01 00:00:00
abstract::Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV), Chronic bee paralysis virus (CBPV), Deformed wing virus (DWV), Sacbrood virus (SBV) and Varroa destructor virus 1 (VDV1) are the six main honeybee viruses reported in Europe. We assessed the accuracy (trueness and precision) of reverse transcriptase quan...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2019.04.020
更新日期:2019-08-01 00:00:00
abstract::Rapid diagnosis of herpes simplex encephalitis (HSE) can only be achieved by the polymerase chain reaction (PCR). In order to carry out PCR under routine conditions, it is of great importance to establish an easy DNA extraction protocol and especially a rapid and sensitive DNA detection method. In the present study, t...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(94)90174-0
更新日期:1994-12-01 00:00:00
abstract::Cell membranes were prepared from herpes simplex virus (HSV) type 1-infected cells and solubilized with a low concentration of sodium deoxycholate. The supernatant after ultracentrifugation was used as antigen in a newly developed solid phase assay, diffusion in gel-enzyme-linked immunosorbent assay (DIG-ELISA). Antig...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(82)90045-3
更新日期:1982-04-01 00:00:00
abstract::Hepatitis B virus is a worldwide public health problem. A simple and effective test to identify viral genotypes would greatly aid efforts to understand and control the spread of this disease. A serial invasive signal amplification reaction assay (PCR-Invader assay) was developed for distinguishing the known eight geno...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.07.014
更新日期:2006-12-01 00:00:00
abstract::Adequate treatment of influenza requires identification of viral type as well as detection of mutation(s) conferring drug resistance. Reverse hybridization-based line probe assays (LiPA) can be performed using several probes immobilized on nitrocellulose, strips enabling LiPA to determine simultaneously viral subtypes...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2012.06.018
更新日期:2012-11-01 00:00:00
abstract::Mungbean yellow mosaic virus-Vigna (MYMV) sequences cloned as partial dimers within the T-DNA of a binary vector were deleted at a high frequency upon conjugal mobilization from Escherichia coli into Agrobacterium tumefaciens. This deletion involving the genome-length viral DNA did not occur when the binary plasmid wa...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.06.002
更新日期:2006-10-01 00:00:00
abstract::The Hawaii agent is a Norwalk-like virus of acute gastroenteritis in humans which is antigenically distinct from the prototype Norwalk agent. We established a solid phase sandwich type microtiter enzyme immunoassay (EIA) for Hawaii antigen employing sera and stools from experimentally challenged volunteers as reagents...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(88)90103-6
更新日期:1988-12-01 00:00:00
abstract::The first case of pandemic influenza A/H1N1/2009 infection was reported in Mexico in mid-April, 2009, and to date the new H1N1 virus has spread to over 160 countries. Therefore, it is important to obtain reliable epidemiological data on the spread of this virus by novel molecular methods can be developed in detection ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.11.033
更新日期:2010-05-01 00:00:00
abstract::In ovo vaccination remains an attractive option for a cost effective, uniform and mass application of vaccines for commercial poultry. However, the vaccines which can be delivered safely by this method are limited and there is no currently licensed embryo-safe vaccine against infectious bronchitis virus (IBV). In this...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.02.019
更新日期:2010-06-01 00:00:00
abstract::Hepatitis A virus (HAV) infection is the leading worldwide cause of acute viral hepatitis. An important aspect of viral control is rapid diagnosis. Epidemiological studies have linked hepatitis A outbreaks to the consumption of drinking water or soft fruits exposed to faecal contamination. Real-time reverse transcript...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.02.017
更新日期:2010-06-01 00:00:00
abstract::Citrus Tristeza Virus (CTV) exists as a large number of distinct strains differing in biological properties and with different distributions in citrus producing countries. Strategies such as eradication or cross protection, aimed at controlling severe variants of the pathogen, require procedures to identify virus stra...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(93)90065-y
更新日期:1993-10-01 00:00:00
abstract::The latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is one of two postulated viral oncogenic proteins. Sequence variations, and in particular a 30 base pair deletion variant called CAO, may define different disease populations. We developed a panel of rat monoclonal antibodies (MAb) specific for the non-w...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2003.10.015
更新日期:2004-03-01 00:00:00
abstract::Performances of serological parallel and serial testing algorithms were analyzed using a combination of three ELISA and three rapid tests for the confirmation of HIV infection. Each was assessed individually for their sensitivity and specificity on a blinded panel of 769 retrospective sera of known HIV status. Western...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.05.021
更新日期:2011-09-01 00:00:00
abstract::Early and accurate diagnosis of Crimean-Congo hemorrhagic fever (CCHF) is essential for the treatment and outcome of the disease and prevention of its further transmission. Molecular-based diagnostic assays now serve as the front-line tool in the diagnosis of CCHF. However, the development of real-time RT-PCR assay fo...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.11.006
更新日期:2006-05-01 00:00:00
abstract::A monoclonal antibody (Mab)-based blocking ELISA was developed for the detection of serum neutralizing antibodies to porcine circovirus type 2 (PCV2). The Mab with neutralizing activity, which was produced by immunizing a recombinant capsid protein of PCV2 expressed in insect cells, was used as the detector antibody. ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.09.023
更新日期:2011-01-01 00:00:00
abstract::Conventional tests for antibody to Hepatitis C virus (HCV) and HCV RNA require considerable time before results are available, remain very expensive and are not adapted to many sub-Saharan African countries where HCV is endemic. The aim of this study was to evaluate the accuracy of an algorithm consisting of two HCV r...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.11.001
更新日期:2006-05-01 00:00:00
abstract::A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of abalone herpesvirus DNA. Two pairs of primers were designed, based on the sequence of the DNA polymerase gene of abalone herpesvirus. The reaction temperature and time were optimized to 63°C and 60min, respectively. LAMP amplicons...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.11.011
更新日期:2014-02-01 00:00:00
abstract::Genotypic drug resistance testing is routine practice in HIV-1 clinical care. The visual interpretation of sequencing electropherograms is labour-intensive and subject to intra- and inter-assay variability because decisions are based on operators' judgments. In this study the performance of the automatic editing tool ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.05.017
更新日期:2013-10-01 00:00:00
abstract::Immunosorbent assays are commonly employed as diagnostic tests in human healthcare, veterinary medicine and bioterrorism prevention. These assays, however, often require long incubation times, limiting sample throughput. As an approach to overcome this weakness, this paper examines the use of rotating capture substrat...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.08.011
更新日期:2006-12-01 00:00:00
abstract::Arenaviruses merit significant attention both as causative agents of endemic hemorrhagic fevers and as model systems to study the immune response to acute and persistent viral infections. Development of highly sensitive quantitative screening methods to detect arenavirus is critical for early diagnosis of patients, to...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.04.001
更新日期:2009-08-01 00:00:00
abstract::A spot nested RT-PCR-RFLP method to detect and identify all members of the Tobamovirus genus is described. It involves a one-step RT-PCR, in which the combination of degenerate deoxyinosine (dI)-substituted primers amplified part of the polymerase region of tobamoviruses, followed by a nested PCR amplification that in...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.01.004
更新日期:2004-05-01 00:00:00
abstract::Human coronavirus NL63 was identified in 2004 in the Netherlands. Due to the high prevalence and world-wide distribution of this pathogen, it is essential to develop a sensitive and specific detection assay suitable for use in a routine diagnostic laboratory. Techniques based on PCR or real-time PCR are laborious and ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.04.024
更新日期:2011-07-01 00:00:00